Pharmaceutical Formulations

ABSTRACT

Improved pharmaceutical compositions are provided comprising one or more solubilized HIV protease inhibiting compounds having improved solubility properties in a medium and/or long chain fatty acid, or mixtures thereof, a pharmaceutically acceptable alcohol, and water.

This application is a continuation of U.S. patent application Ser. No.11/546,673, filed Oct. 12, 2006, now pending, which is a continuation ofU.S. patent application Ser. No. 09/576,097, filed May 22, 2000, nowU.S. Pat. No. 7,141,593, which claims the benefit of U.S. ProvisionalApplication No. 60/137,634, filed Jun. 4, 1999, and U.S. ProvisionalApplication No. 60/177,020, filed Jan. 19, 2000. This applicationincorporates by reference U.S. Pat. No. 7,141,593 in its entirety.

TECHNICAL FIELD

This invention relates to improved pharmaceutical formulationscomprising at least one HIV protease inhibiting compound in apharmaceutically acceptable solution of a medium and/or long chain fattyacid, ethanol or propylene glycol, and water, wherein said HIV proteaseinhibiting compound contained therein has improved solubilityproperties.

BACKGROUND OF THE INVENTION

Inhibitors of human immunodeficiency virus (HIV) protease have beenapproved for use in the treatment of HIV infection for several years. Aparticularly effective HIV protease inhibitor is(2S,3S,5S)-5-(N-(N-((N-methyl-N-((2-isopropyl-4-thiazolyl)-methyl)amino)carbonyl)-L-valinyl)amino)-2-(N-((5-thiazolyl)methoxy-carbonyl)-amino)-1,6-diphenyl-3-hydroxyhexane(ritonavir), which is marketed as NORVIR®. Ritonavir is known to haveutility for the inhibition of HIV protease, the inhibition of HIVinfection, and the enhancement of the pharmacokinetics of compoundswhich are metabolized by cytochrome P₄₅₀ monooxygenase. Ritonavir isparticularly effective for the inhibition of HIV infection when usedalone or in combination with one or more reverse transcriptaseinhibitors and/or one or more other HIV protease inhibitors.

HIV protease inhibiting compounds typically are characterized by havingpoor oral bioavailability, and there is a continuing need for thedevelopment of improved oral dosage forms for HIV protease inhibitorshaving suitable oral bioavailability, stability, and side effectsprofiles.

Ritonavir and processes for its preparation are disclosed in U.S. Pat.No. 5,541,206, issued Jul. 30, 1996, the disclosure of which is hereinincorporated by reference. This patent discloses processes for preparingritonavir which produce a crystalline polymorph of ritonavir, known ascrystalline Form I.

Another process for the preparation of ritonavir is disclosed in U.S.Pat. No. 5,567,823, issued Oct. 22, 1996, the disclosure of which isherein incorporated by reference. The process disclosed in this patentalso produces ritonavir as crystalline Form I.

Pharmaceutical compositions comprising ritonavir or a pharmaceuticallyacceptable salt thereof are disclosed in U.S. Pat. Nos. 5,541,206,issued Jul. 30, 1996; 5,484,801, issued Jan. 16, 1996; 5,725,878, issuedMar. 10, 1998; and 5,559,158, issued Sep. 24, 1996 and in InternationalApplication No. WO98/22106, published May 28, 1998 (corresponding toU.S. Ser. No. 08/966,495, filed Nov. 7, 1997), the disclosures of all ofwhich are herein incorporated by reference.

The use of ritonavir to inhibit an HIV infection is disclosed in U.S.Pat. No. 5,541,206, issued Jul. 30, 1996. The use of ritonavir incombination with one or more reverse transcriptase inhibitors to inhibitan HIV infection is disclosed in U.S. Pat. No. 5,635,523, issued Jun. 3,1997. The use of ritonavir in combination with one or more HIV proteaseinhibitors to inhibit an HIV infection is disclosed in U.S. Pat. No.5,674,882, issued Oct. 7, 1997. The use of ritonavir to enhance thepharmacokinetics of compounds metabolized by cytochrome P450monooxygenase is disclosed in WO 97/01349, published Jan. 16, 1997(corresponding to U.S. Ser. No. 08/687,774, filed Jun. 26, 1996). Thedisclosures of all of these patents and patent applications are hereinincorporated by reference.

Examples of HIV protease inhibiting compounds include:

N-(2(R)-hydroxy-1(S)-indanyl)-2(R)-phenylmethyl-4(S)-hydroxy-5-(1-(4-(3-pyridylmethyl)-2(S)-N′-(t-butylcarboxamido)-piperazinyl))-pentaneamide (for example, indinavir) and related compounds,disclosed in European Patent Application No. EP 541168, published May12, 1993, and U.S. Pat. No. 5,413,999, issued May 9, 1995, both of whichare herein incorporated by reference;N-tert-butyl-decahydro-2-[2(R)-hydroxy-4-phenyl-3(S)-[[N-(2-quinolylcarbonyl)-L-asparaginyl]amino]butyl]-(4aS,8aS)-isoquinoline-3(S)-carboxamide(for example, saquinavir) and related compounds, disclosed in U.S. Pat.No. 5,196,438, issued Mar. 23, 1993, which is incorporated herein byreference;5(S)-Boc-amino-4(S)-hydroxy-6-phenyl-2(R)-phenylmethylhexanoyl-(L)-Val-(L)-Phe-morpholin-4-ylamideand related compounds, disclosed in European Patent Application No.EP532466, published Mar. 17, 1993, which is incorporated herein byreference;1-Naphthoxyacetyl-beta-methylthio-Ala-(2S,3S)-3-amino-2-hydroxy-4-butanoyl1,3-thiazolidine-4-t-butylamide (for example,1-Naphthoxyacetyl-Mta-(2S,3S)-AHPBA-Thz-NH-tBu),5-isoquinolinoxyacetyl-beta-methylthio-Ala-(2S,3S)-3-amino-2-hydroxy-4-butanoyl-1,3-thiazolidine-4-t-butylamide,and related compounds, disclosed in European Patent Application No.EP490667, published Jun. 17, 1992 and Chem. Pharm. Bull. 40 (8) 2251(1992), which are both incorporated herein by reference;[1 S-[1R-(R-),2S*])-N¹[3-[[[(1,1-dimethylethyl)amino]carbonyl](2-methylpropyl)amino]-2-hydroxy-1-(phenylmethyl)propyl]-2-[(2-quinolinylcarbonyl)amino]-butanediamide(for example, SC-52151) and related compounds, disclosed in PCT PatentApplication No. WO92/08701, published May 29, 1992 and PCT PatentApplication No. WO93/23368, published Nov. 25, 1993, both of which areherein incorporated by reference;

(for example, VX-478) and related compounds, disclosed in PCT PatentApplication No. WO 94/05639, published Mar. 17, 1994, which isincorporated herein by reference;

(for example, DMP-323) or

(for example, DMP-450) and related compounds, disclosed in PCT PatentApplication No. W0 93/07128, published Apr. 15, 1993, which isincorporated herein by reference;

(for example, AG1343, (nelfinavir)), disclosed in PCT Patent ApplicationNo. W0 95/09843, published Apr. 13, 1995 and U.S. Pat. No. 5,484,926,issued Jan. 16, 1996, which are both incorporated herein by reference;

(for example, BMS 186,318) disclosed in European Patent Application No.EP580402, published Jan. 26, 1994, which is incorporated herein byreference;

(for example, SC-55389a) and related compounds disclosed in PCT PatentApplication No. WO 9506061, published Mar. 2, 1995, which isincorporated herein by reference and at 2nd National Conference on HumanRetroviruses and Related Infections, (Washington, D.C., Jan. 29-Feb. 2,1995), Session 88;

(for example, BILA 1096 BS) and related compounds disclosed in EuropeanPatent Application No. EP560268, published Sep. 15, 1993, which isincorporated herein by reference; and

(for example, U-140690 (tipranavir)) and related compounds disclosed inPCT Patent Application No. WO 9530670, published Nov. 16, 1995, and U.S.Pat. No. 5,852,195, issued Dec. 22, 1998, the disclosures of both ofwhich are herein incorporated by reference; or a pharmaceuticallyacceptable salt of any of the above.

Another example of an HIV protease inhibiting compound includes acompound of formula I:

or a pharmaceutically acceptable salt thereof, disclosed in PCT PatentApplication No. W0 94/14436, published Jul. 7, 1994, and U.S. Pat. No.5,541,206, issued Jul. 30, 1996, the disclosures of both of which areherein incorporated by reference.

The compounds of formula I are useful to inhibit HIV infections and,thus, are useful for the treatment of AIDS.

Another example of an HIV protease inhibiting compound is a compound offormula II:

and related compounds, or a pharmaceutically-acceptable salt thereof, asdisclosed in U.S. patent application Ser. No. 08/572,226, filed Dec. 13,1996 and U.S. patent application Ser. No. 08/753,201, filed Nov. 21,1996, and International Patent Application No. WO 97/21685, publishedJun. 19, 1997, the disclosures of which are herein incorporated byreference. A preferred compound of formula II is known as ABT-378 andhas a chemical name of(2S,3S,5S)-2-(2,6-dimethylphenoxyacetyl)-amino-3-hydroxy-5-(2S-(1-tetrahydropyrimid-2-onyl)-3-methyl-butanoyl)amino-1,6-diphenylhexane,or a pharmaceutically-acceptable salt thereof. The preparation of thiscompound is disclosed in U.S. Pat. No. 5,914,332, issued Jun. 22, 1999,the disclosure of which is herein incorporated by reference.

Solubility is an important factor in the formulation of HIV proteaseinhibiting compounds. Compounds of formula I typically have an aqueoussolubility of approximately 6 micrograms per milliliter at pH>2. This isconsidered to be extremely poor aqueous solubility and, therefore, acompound of formula I in the free base form would be expected to providevery low oral bioavailability. In fact, the free base form of a compoundof formula I, administered as an unformulated solid in a capsule dosageform, is characterized by a bioavailability of less than 2% following a5 mg/kg oral dose in dogs.

Acid addition salts of a compound of formula I (for example,bishydrochloride, bis-tosylate, bis-methane sulfonate and the like) haveaqueous solubilities of <0.1 milligrams/milliliter. This is only aslight improvement over the solubility of the free base. This lowaqueous solubility would not make practical the administration oftherapeutic amounts of an acid addition salt of a compound of formula Ias an aqueous solution. Furthermore, in view of this low aqueoussolubility, it is not surprising that the bis-tosylate of a compound offormula I, administered as an unformulated solid in a capsule dosageform, is characterized by a bioavailability of less than 2% following a5 mg/kg oral dose in dogs.

In order to have a suitable oral dosage form of a compound of formula I,the oral bioavailability of a compound of formula I should be at least20%. Preferably, the oral bioavailability of a compound of formula Ifrom the dosage form should be greater than about 40% and, morepreferably, greater than about 50%.

One measure of the potential usefulness of an oral dosage form of apharmaceutical agent is the bioavailability observed after oraladministration of the dosage form. Various factors can affect thebioavailability of a drug when administered orally. These factorsinclude aqueous solubility, drug absorption, dosage strength and firstpass effect. Aqueous solubility is one of the most important of thesefactors. When a drug has poor aqueous solubility, attempts are oftenmade to identify salts or other derivatives of the drug which haveimproved aqueous solubility. When a salt or other derivative of the drugis identified which has good aqueous solubility, it is generallyaccepted that an aqueous solution formulation of this salt or derivativewill provide the optimum oral bioavailability. The bioavailability ofthe oral solution formulation of a drug is then generally used as thestandard bioavailability against which other oral dosage forms can bemeasured.

For a variety of reasons, such as patient compliance and taste masking,a solid dosage form, such as capsules, is usually preferred over aliquid dosage form. However, oral solid dosage forms, such as a tabletor a powder, and the like, of a drug generally provide a lowerbioavailability than oral solutions of the drug. One goal of thedevelopment of a suitable capsule dosage form is to obtain abioavailability of the drug that is as close as possible to thebioavailability demonstrated by the oral solution formulation of thedrug.

While some drugs would be expected to have good solubility in organicsolvents, it would not necessarily follow that oral administration ofsuch a solution would give good bioavailability for the drug. It hasbeen found that a compound of formula I has good solubility inpharmaceutically acceptable organic solvents and that the solubility insuch solvents is enhanced in the presence of a pharmaceuticallyacceptable long chain fatty acid. Administration of the solution as anencapsulated dosage form (soft elastic capsules or hard gelatincapsules) provides an oral bioavailability of as high as about 60% ormore.

Thus, it would be an important contribution to the art to provide animproved pharmaceutical formulation comprising at least one solubilizedHIV protease inhibiting compound having enhanced solubility properties.

In addition, the administration of ritonavir and a compound which ismetabolized by cytochrome P450 monooxygenase is useful for improving inhumans the pharmacokinetics of the compound which is metabolized bycytochrome P450 monooxygenase.

A method of improving the pharmacokinetics of a drug (or apharmaceutically acceptable salt thereof) which is metabolized bycytochrome P450 monooxygenase comprises coadministering ritonavir or apharmaceutically acceptable salt thereof. When administered incombination, the two therapeutic agents can be formulated as separatecompositions which are administered at the same time or different times,or the two therapeutic agents can be administered as a singlecomposition.

Drugs which are metabolized by cytochrome P450 monooxygenase and whichbenefit from coadministration with ritonavir include theimmunosuppressants cyclosporine, FK-506 and rapamycin, thechemotherapeutic agents taxol and taxotere, the antibioticclarithromycin and the HIV protease inhibitors A-77003, A-80987, MK-639,saquinavir, VX-478, AG1343, DMP-323, XM-450, BILA 2011 BS, BILA 1096 BS,BILA 2185 BS, BMS 186,318, LB71262, SC-52151, SC-629(N,N-dimethylglycyl-N-(2-hyrdoxy-3-(((4-methoxyphenyl)sulphonyl)(2-methylpropyl)amino)-1-(phenylmethyl)propyl)-3-methyl-L-valinamide),KNI-272, CGP 53437, CGP 57813 and U-103017.

A method for improving the pharmacokinetics of an HIV protease inhibitor(or a pharmaceutically acceptable salt thereof) which is metabolized bycytochrome P450 monooxygenase comprises coadministering ritonavir or apharmaceutically acceptable salt thereof. Such a combination ofritonavir or a pharmaceutically acceptable salt thereof and an HIVprotease inhibitor or a pharmaceutically acceptable salt thereof whichis metabolized by cytochrome P450 monooxygenase is useful for inhibitingHIV protease in humans and is also useful for inhibition, treatment orprophylaxis of an HIV infection or AIDS (acquired immune deficiencysyndrome) in humans. When administered in combination, the twotherapeutic agents can be formulated as separate compositions which areadministered at the same time or different times, or the two therapeuticagents can be administered as a single composition.

The total daily dose of ritonavir to be administered to a human or othermammal host in single or divided doses may be in amounts, for example,from 0.001 to 300 mg/kg body weight daily and more usually 0.1 to 50mg/kg and even more usually 0.1 to 25 mg/kg. Dosage unit compositionsmay contain such amounts of submultiples thereof to make up the dailydose.

The total daily dose of the drug which is metabolized by cytochrome P450monooxygenase to be administered to a human or other mammal is wellknown and can be readily determined by one of ordinary skill in the art.Dosage unit compositions may contain such amounts of submultiplesthereof to make up the daily dose.

Liquid dosage forms for oral administration may include pharmaceuticallyacceptable emulsions, solutions, suspensions, syrups, and elixirscontaining inert diluents commonly used in the art, such as water. Suchcompositions may also comprise adjuvants, such as wetting agents,emulsifying and suspending agents, and sweetening, flavoring, andperfuming agents.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates the powder X-ray diffraction pattern of thesubstantially pure Form I crystalline polymorph of ritonavir.

FIG. 2 illustrates the powder X-ray diffraction pattern of thesubstantially pure Form II crystalline polymorph of ritonavir.

FIG. 3 illustrates the equilibrium solubility of Ritonavir Form II inthe premix provided in Example 9.

FIG. 4 illustrates the equilibrium solubility of Ritonavir Form I in thepremix provided in Example 9.

FIG. 5 illustrates the effect of added water on the solubility ofRitonavir Form II in oleic acid+ethanol co-solvent system.

FIG. 6 illustrates the dissolution profile of Ritonavir Form II crystalsin the premix provided in Example 9.

FIG. 7 illustrates the 3D plots for the solubility of Ritonavir Form Iand II as a function of temperature, water, and ethanol in the premixprovided in Example 9.

SUMMARY OF THE INVENTION

The instant invention provides pharmaceutical compositions comprising atleast one solubilized HIV protease inhibiting compound in apharmaceutically acceptable solution of medium and/or long chain fattyacids or mixtures thereof, a pharmaceutically acceptable alcohol, andwater, wherein said solubilized HIV protease inhibiting compoundscontained therein have improved solubility properties.

DETAILED DESCRIPTION OF THE INVENTION

The instant invention comprises a solubilized HIV protease inhibitingcompound or a combination of solubilized HIV protease inhibitingcompounds, or pharmaceutically acceptable salts thereof, in apharmaceutically acceptable organic solvent comprising a mixture of atleast one pharmaceutically acceptable medium and/or long chain fattyacid, a pharmaceutically-acceptable alcohol, and water.

The compositions of the instant invention provide greatly improvedsolubility for said solubilized HIV protease inhibiting compoundscontained therein when compared to analogous compositions without theaddition of water.

A preferred composition of the invention is a solution comprising (a) asolubilized HIV protease inhibiting compound or a combination ofsolubilized HIV protease inhibiting compounds (preferably, a compound ofthe formula I or II, or saquinavir or nelfinavir or indinavir or, morepreferably, ritonavir or ABT-378 or saquinavir or nelfinavir orindinavir, or, most preferably, ritonavir or ABT-378); or a combinationof ritonavir or nelfinavir and another HIV protease inhibitor(preferably, ABT-378 or saquinavir or indinavir or nelfinavir, or, morepreferably, a combination of ritonavir or nelfinavir and another HIVprotease inhibitor (preferably, ABT-378 or saquinavir or indinavir ornelfinavir), or, most preferably, a combination of ritonavir andABT-378) in the amount of from about 1% to about 50% (preferably, fromabout 1% to about 40%; more preferably, from about 10% to about 40% byweight of the total solution,

(b) a pharmaceutically acceptable organic solvent which comprises (i) apharmaceutically acceptable medium and/or long chain fatty acid ormixtures thereof in the amount of from about 20% to about 99%(preferably, from about 30% to about 75% by weight of the total solutionor (ii) a mixture of (1) a pharmaceutically acceptable medium and/orlong chain fatty acid or mixtures thereof in the amount of from about20% to about 99% (preferably, from about 30% to about 75% by weight ofthe total solution; (2) ethanol in the amount of from about 1% to about15% (preferably, from about 3% to about 12%) by weight of the totalsolution, or, alternatively, propylene glycol in the amount of fromabout 1% to about 15% (preferably, from about 5% to about 10%); (c)water in the amount of from about 0.4% to about 3.5%; and optionally,(d) a pharmaceutically acceptable surfactant in the amount of from about0% to about 40% (preferably, from about 2% to about 20% and mostpreferably, from about 2.5% to about 15%) by weight of the totalsolution.

In a preferred embodiment of the invention, the solution is encapsulatedin a soft elastic gelatin capsule (SEC) or a hard gelatin capsule, ororally ingested after further dilution in an appropriate diluent orvehicle.

Specifically, preferred ratios (w/w) of ritonavir to ABT-378 are fromabout 1:16 to about 5:1. Even more preferred is a ratio of ritonavir toABT-378 of from about 1:8 to about 3:1. An even more preferred ratio ofritonavir to ABT-378 is 1:4.

Solutions as described herein may include micellar solutions, which arethermodynamically stable systems formed spontaneously in water above acritical temperature and concentration. Said micellar solutions containsmall colloidal aggregates (micelles), the molecules of which are inrapid thermodynamic equilibrium with a measurable concentration ofmonomers. Micellar solutions exhibit solubilization phenomena andthermodynamic stability.

Preferably, the pharmaceutically acceptable organic solvent comprisesfrom about 50% to about 99% by weight of the total solution. Morepreferably, the pharmaceutically acceptable organic solvent or mixtureof pharmaceutically acceptable organic solvents comprises from about 50%to about 75% by weight of the total solution.

The term “pharmaceutically acceptable medium and/or long chain fattyacid” as used herein refers to saturated or unsaturated C₈ to C₂₄ fattyacids. Preferred fatty acids are mono-unsaturated C₁₆-C₂₀ fatty acidswhich are liquids at room temperature. A most preferred fatty acid isoleic acid, with or without additional medium and/or long chain fattyacids in the mixture. One suitable source of said oleic acid is HenkelCorporation.

The term “pharmaceutically acceptable alcohol” as used herein refers toalcohols which are liquid at room temperature, for example ethanol,propylene glycol, 2-2(ethoxyethoxy)ethanol (Transcutol®, Gattefosse,Westwood, N.J.), benzyl alcohol, glycerol, polyethylene glycol 200,polyethylene glycol 300, polyethylene glycol 400, and the like, ormixtures thereof.

Preferred pharmaceutically acceptable solvents comprise (1)pharmaceutically acceptable medium and/or long chain fatty acid in theamount of from about 40% to about 75% by weight of the total solution;(2) ethanol or propylene glycol in the amount of from about 1% to about15% by weight of the total solution; and (3) water in the amount of fromabout 0.4% to about 3.5% by weight of the total solution. More preferredpharmaceutically acceptable solvents comprise (1) a pharmaceuticallyacceptable medium and/or long chain fatty acid in the amount of fromabout 40% to about 75% by weight of the total solution and (2) ethanolor propylene glycol in the amount of from about 3% to about 12% byweight of the total solution. Even more preferred pharmaceuticallyacceptable solvents comprise (1) oleic acid in the amount of from about40% to about 75% by weight of the total solution and (2) ethanol orpropylene glycol in the amount of from about 3% to about 12% by weightof the total solution.

In one embodiment of the invention, a more preferred composition of theinvention is a solution comprising (a) solubilized ritonavir in theamount of from about 1% to about 30% (preferably, from about 5% to about25%) by weight of the total solution, (b) a pharmaceutically acceptableorganic solvent which comprises (i) a pharmaceutically acceptable mediumand/or long chain fatty acid in the amount of from about 40% to about99% (preferably, from about 30% to about 75% by weight of the totalsolution or (ii) a mixture of (1) a pharmaceutically acceptable longchain fatty acid in the amount of from about 40% to about 99%(preferably, from about 30% to about 75% by weight of the total solutionand (2) ethanol in the amount of from about 1% to about 15% (preferably,from about 3% to about 12%) by weight of the total solution, (c) waterin the amount of from about 0.4% to about 3.5% and (d) apharmaceutically acceptable surfactant in the amount of from about 0% toabout 20% (preferably, from about 2.5% to about 10%) by weight of thetotal solution.

In a more preferred embodiment of the invention, the solution isencapsulated in a soft elastic gelatin capsule (SEC) or a hard gelatincapsule.

An even more preferred composition of the invention is a solutioncomprising (a) solubilized ritonavir in the amount of from about 1% toabout 30% (preferably, from about 5% to about 25%) by weight of thetotal solution, (b) a pharmaceutically acceptable organic solvent whichcomprises (i) oleic acid in the amount of from about 15% to about 99%(preferably, from about 30% to about 75% by weight of the total solutionor (ii) a mixture of (1) oleic acid in the amount of from about 15% toabout 99% (preferably, from about 30% to about 75% by weight of thetotal solution and (2) ethanol in the amount of from about 1% to about15% (preferably, from about 3% to about 12%) by weight of the totalsolution, (c) water in the amount of from about 0.4% to about 3.5%, and(d) polyoxyl 35 castor oil in the amount of from about 0% to about 20%(preferably, from about 2.5% to about 10%) by weight of the totalsolution.

In an even more preferred embodiment of the invention, the solution isencapsulated in a soft elastic gelatin capsule (SEC) or a hard gelatincapsule.

A most preferred composition of the invention is a solution comprising(a) solubilized ritonavir in the amount of about 10% by weight of thetotal solution, (b) a pharmaceutically acceptable organic solvent whichcomprises a mixture of (1) oleic acid in the amount of from about 70% toabout 75% by weight of the total solution and (2) ethanol in the amountof from about 3% to about 12%, preferably, about 12%, by weight of thetotal solution, (c) water in the amount of from about 0.4% to about 1.5%and (d) polyoxyl 35 castor oil in the amount of about 6% by weight ofthe total solution.

In a most preferred embodiment of the invention, the solution isencapsulated in a soft elastic gelatin capsule (SEC) or a hard gelatincapsule and the solution also comprises an antioxidant (preferably, BHT(butylated hydroxytoluene)) in the amount of about 0.025% by weight ofthe total solution.

In one embodiment of the invention, a more preferred composition of theinvention is a solution comprising (a) a combination of solubilized HIVprotease inhibiting compounds which are ritonavir and ABT-378 in theamount of from about 1% to about 45% (preferably, from about 5% to about45%) by weight of the total solution, and (b) a pharmaceuticallyacceptable organic solvent which comprises (i) a pharmaceuticallyacceptable medium and/or long chain fatty acid in the amount of fromabout 40% to about 99% (preferably, from about 30% to about 75% byweight of the total solution or (ii) a mixture of (1) a pharmaceuticallyacceptable long chain fatty acid in the amount of from about 40% toabout 99% (preferably, from about 30% to about 75% by weight of thetotal solution and (2) propylene glycol in the amount of from about 1%to about 15% by weight of the total solution, (c) water in the amount offrom about 0.4% to about 3.5% and (d) a pharmaceutically acceptablesurfactant in the amount of from about 0% to about 20% (preferably, fromabout 2.5% to about 10%) by weight of the total solution.

In a more preferred embodiment of the invention, the solution isencapsulated in a soft elastic gelatin capsule (SEC) or a hard gelatincapsule.

An even more preferred composition of the invention is a solutioncomprising (a) a combination of solubilized HIV protease inhibitingcompounds which are ritonavir and ABT-378 in the amount of from about 1%to about 45% (preferably, from about 5% to about 45%) by weight of thetotal solution, (b) a pharmaceutically acceptable organic solvent whichcomprises (i) oleic acid in the amount of from about 15% to about 99%(preferably, from about 30% to about 75% by weight of the total solutionor (ii) a mixture of (1) oleic acid in the amount of from about 15% toabout 99% (preferably, from about 30% to about 75% by weight of thetotal solution and (2) propylene glycol in the amount of from about 1%to about 8% by weight of the total solution, (c) water in the amount offrom about 0.4% to about 3.5%, and (d) polyoxyl 35 castor oil in theamount of from about 0% to about 20% (preferably, from about 2.5% toabout 10%) by weight of the total solution.

In an even more preferred embodiment of the invention, the solution isencapsulated in a soft elastic gelatin capsule (SEC) or a hard gelatincapsule.

A most preferred composition of the invention is a solution comprising(a) a combination of solubilized HIV protease inhibiting compounds whichare ritonavir and ABT-378 in the amount of about 10% by weight of thetotal solution, (b) a pharmaceutically acceptable organic solvent whichcomprises a mixture of (1) oleic acid in the amount of from about 70% toabout 75% by weight of the total solution and (2) propylene glycol inthe amount of from about 1% to about 15%, preferably, about 6%, byweight of the total solution, (c) water in the amount of from about 0.4%to about 1.5% and (d) polyoxyl 35 castor oil in the amount of about 6%by weight of the total solution.

In a most preferred embodiment of the invention, the solution isencapsulated in a soft elastic gelatin capsule (SEC) or a hard gelatincapsule and the solution also comprises an antioxidant (preferably, BHT(butylated hydroxytoluene)) in the amount of about 0.025% by weight ofthe total solution.

The amount of water employed in the pharmaceutical composition of theinstant invention comprises from about 0.4% to about 3.5% by weight ofthe total solution of water. Preferably, the weight of the totalsolution of water is from about 0.4% to about 2.0%; more preferably fromabout 0.4% to about 1.5%; and the most preferred being about 1%.

In addition, the composition of the invention can comprise antioxidants(for example, ascorbic acid, BHA (butylated hydroxyanisole), BHT(butylated hydroxytoluene), vitamin E, and the like) for chemicalstability.

The term “pharmaceutically acceptable acid” as used herein refers to (i)an inorganic acid such as hydrochloric acid, hydrobromic acid,hydroiodic acid and the like, (ii) an organic mono-, di- ortri-carboxylic acid (for example, formic acid, acetic acid, adipic acid,alginic acid, citric acid, ascorbic acid, aspartic acid, benzoic acid,butyric acid, camphoric acid, gluconic acid, glucuronic acid,galactaronic acid, glutamic acid, heptanoic acid, hexanoic acid, fumaricacid, lactic acid, lactobionic acid, malonic acid, maleic acid,nicotinic acid, oxalic acid, pamoic acid, pectinic acid,3-phenylpropionic acid, picric acid, pivalic acid, propionic acid,succinic acid, tartaric acid, undecanoic acid and the like) or (iii) asulfonic acid (for example, benzenesulfonic acid, sodium bisulfate,sulfuric acid, camphorsulfonic acid, dodecylsulfonic acid,ethanesulfonic acid, methanesulfonic acid, isethionic acid,naphthalenesulfonic acid, p-toluenesulfonic acid and the like).

The term “pharmaceutically acceptable surfactant” as used herein refersto a pharmaceutically acceptable non-ionic surfactant for example,polyoxyethylene castor oil derivatives (for example,polyoxyethyleneglyceroltriricinoleate or polyoxyl ethylene 35 castor oil(Cremophor® EL, BASF Corp.) or polyoxyethyleneglycerol oxystearate(Cremophor® RH 40 (glycerol polyethyleneglycol oxystearate) orCremophor® RH 60 (polyethyleneglycol 60 hydrogenated castor oil), BASFCorp., and the like) or block copolymers of ethylene oxide and propyleneoxide, also known as polyoxyethylene polyoxypropylene block copolymersor polyoxyethylenepolypropylene glycol, such as Poloxamer®124,Poloxamer® 188, Poloxamer® 237, Poloxamer® 338, Poloxamer® 407, and thelike, (BASF Wyandotte Corp.) or a mono fatty acid ester ofpolyoxyethylene (20) sorbitan (for example, polyoxyethylene (20)sorbitan monooleate (Tween® 80), polyoxyethylene (20) sorbitanmonostearate (Tween® 60), polyoxyethylene (20) sorbitan monopalmitate(Tween® 40), polyoxyethylene (20) sorbitan monolaurate (Tweens® 20)) andthe like) or a sorbitan fatty acid ester (including sorbitan laurate,sorbitan oleate, sorbitan palmitate, sorbitan stearate and the like). Apreferred pharmaceutically acceptable surfactant is polyoxyl 35 castoroil (Cremophor® EL, BASF Corp.), polyoxyethylene (20) sorbitanmonolaurate (Tween®) 20), polyoxyethylene (20) sorbitan monooleate(Tween® 80) or a sorbitan fatty acid ester, for example sorbitan oleate.A most preferred pharmaceutically acceptable surfactant is polyoxyl 35castor oil (Cremophor® EL, BASF Corp.).

As used herein, the term “substantially pure”, when used in reference toa polymorph of ritonavir, refers to a polymorph of ritonavir, Form I orForm II, which is greater than about 90% pure. This means that thepolymorph of ritonavir does not contain more than about 10% of any othercompound and, in particular, does not contain more than about 10% of anyother form of ritonavir. More preferably, the term “substantially pure”refers to a polymorph of ritonavir, Form I or Form II, which is greaterthan about 95% pure. This means that the polymorph of ritonavir does notcontain more than about 5% of any other compound and, in particular,does not contain more than about 5% of any other form of ritonavir. Evenmore preferably, the term “substantially pure” refers to a polymorph ofritonavir, Form I or Form II, which is greater than about 97% pure. Thismeans that the polymorph of ritonavir does not contain more than about3% of any other compound and, in particular, does not contain more thanabout 3% of any other form of ritonavir.

As used herein, the term “substantially pure”, when used in reference toamorphous ritonavir, refers to amorphous ritonavir which is greater thanabout 90% pure. This means that the amorphous ritonavir does not containmore than about 10% of any other compound and, in particular, does notcontain more than about 10% of any other form of ritonavir. Morepreferably, the term “substantially pure”, when used in reference toamorphous ritonavir, refers to amorphous ritonavir, which is greaterthan about 95% pure. This means that the amorphous ritonavir does notcontain more than about 5% of any other compound and, in particular,does not contain more than about 5% of any other form of ritonavir. Evenmore preferably, the term “substantially pure”, when used in referenceto amorphous ritonavir, refers to amorphous ritonavir which is greaterthan about 97% pure. This means that the amorphous ritonavir does notcontain more than about 3% of any other compound and, in particular,does not contain more than about 3% of any other form of ritonavir.

The composition and preparation of soft elastic gelatin capsules is wellknown in the art. The composition of a soft elastic gelatin capsuletypically comprises from about 30% to about 50% by weight of gelatin NF& EP, from about 20% to about 30% by weight of a plasticizer, and fromabout 25% to about 40% by weight of water. Plasticizers useful in thepreparation of soft elastic gelatin capsules are glycerin, sorbitol, orpropylene glycol and the like, or combinations thereof. A preferred softelastic gelatin capsule has a composition comprising gelatin NF & EP(Type 195) (about 42.6% by weight), glycerin (USP) (about 96% active;about 13.2% by weight), purified water (USP) (about 27.4% by weight),sorbitol special (about 16% by weight) and titanium dioxide (USP) (about0.4% by weight).

The soft elastic gelatin capsule material can also comprise additivessuch as preservatives, opacifiers, dyes or flavors, and the like.

Various methods can be used for manufacturing and filling the softelastic gelatin capsules, for example, a seamless capsule method, arotary method (developed by Scherer) or a method using a Liner® machineor an Accogel® machine, and the like. Also various manufacturingmachines can be used for manufacturing the capsules.

Hard gelatin capsules are purchased from Capsugel, Greenwood, S.C.Capsules are filled manually or by capsule filling machine. The targetfilling volume/weight depends on the potency of the filling solution incombination with the desired dosage strength.

In general, the compositions of this invention can be prepared in thefollowing manner. The pharmaceutically acceptable medium and/or longchain fatty acid and ethanol or propylene glycol and water are mixed ata temperature from 15-30° C., along with the antioxidant. The HIVprotease inhibitor, or mixture of HIV protease inhibitors, is added andstirred until dissolved. The pharmaceutically acceptable surfactant isadded with mixing. The appropriate volume of the resulting mixtureneeded to provide the desired dose of the HIV protease inhibitingcompound(s) is filled into hard gelatin capsules or soft elastic gelatincapsules.

Similar increases in the solubility of HIV protease inhibitors in oralsolution formulations may be obtained by the addition of water in rangesas disclosed herein. Oral solution formulations are disclosed in U.S.Pat. No. 5,484,801, issued Jan. 16, 1996, the disclosure of which isherein incorporated by reference.

EXAMPLES

The following Examples will serve to further illustrate the instantinvention.

Powder X-ray diffraction analysis of samples was conducted in thefollowing manner. Samples for X-ray diffraction analysis were preparedby spreading the sample powder (with no prior grinding required) in athin layer on the sample holder and gently flattening the sample with amicroscope slide.

A Nicolet 12/V X-ray Diffraction System was used with the followingparameters: X-ray source: Cu-Kα1; Range: 2.00-40.00° Two Theta; ScanRate: 1.00 degree/minute; Step Size: 0.02 degrees; Wavelength: 1.540562angstroms.

Characteristic powder X-ray diffraction pattern peak positions arereported for polymorphs in terms of the angular positions (two theta)with an allowable variability of ±0.1°. This allowable variability isspecified by the U.S. Pharmacopeia, pages 1843-1844 (1995). Thevariability of +0.10 is intended to be used when comparing two powderX-ray diffraction patterns. In practice, if a diffraction pattern peakfrom one pattern is assigned a range of angular positions (two theta)which is the measured peak position±0.1° and a diffraction pattern peakfrom the other pattern is assigned a range of angular positions (twotheta) which is the measured peak position±0.1° and if those ranges ofpeak positions overlap, then the two peaks are considered to have thesame angular position (two theta). For example, if a diffraction patternpeak from one pattern is determined to have a peak position of 5.20°,for comparison purposes the allowable variability allows the peak to beassigned a position in the range of 5.10°-5.30°. If a comparison peakfrom the other diffraction pattern is determined to have a peak positionof 5.35°, for comparison purposes the allowable variability allows thepeak to be assigned a position in the range of 5.25°-5.45°. Becausethere is overlap between the two ranges of peak positions (for example,5.10°-5.30° and 5.25°-5.45°) the two peaks being compared are consideredto have the same angular position (two theta).

Solid state nuclear magnetic resonance analysis of samples was conductedin the following manner. A Bruker AMX-400 MHz instrument was used withthe following parameters: CP-MAS (cross-polarized magic angle spinning);spectrometer frequency for 13C was 100.627952576 MHz; pulse sequence wascp21ev; contact time was 2.5 milliseconds; temperature was 27.0° C.;spin rate was 7000 Hz; relaxation delay was 6.000 sec; 1st pulse widthwas 3.8 microseconds; 2nd pulse width was 8.6 microseconds; acquisitiontime was 0.034 seconds; sweep width was 30303.0 Hz; 2000 scans.

FT near infrared analysis of samples was conducted in the followingmanner. Samples were analyzed as neat, undiluted powders contained in aclear glass 1 dram vial. A Nicolet Magna System 750 FT-IR spectrometerwith a Nicolet SabIR near infrared fiber optic probe accessory was usedwith the following parameters: the source was white light; the detectorwas PbS; the beamsplitter was CaF2; sample spacing was 1.0000; digitizerbits was 20; mirror velocity was 0.3165; the aperture was 50.00; samplegain was 1.0; the high pass filter was 200.0000; the low pass filter was11000.0000; the number of sample scans was 64; the collection length was75.9 seconds; the resolution was 8.000; the number of scan points was8480; the number of FFT points was 8192; the laser frequency was 15798.0cm⁻¹; the interferogram peak position was 4096; the apodization wasHapp-Genzel; the number of background scans was 64 and the backgroundgain was 1.0.

FT mid infrared analysis of samples was conducted in the followingmanner. Samples were analyzed as neat, undiluted powders. A NicoletMagna System 750 FT-IR spectrometer with a Spectra-Tech InspectIR videomicroanalysis accessory and a Germanium attenuated total reflectance (GeATR) crystal was used with the following parameters: the source wasinfrared; the detector was MCT/A; the beamsplitter was KBr; samplespacing was 2.0000; digitizer bits was 20; mirror velocity was 1.8988;the aperture was 100.00; sample gain was 1.0; the high pass filter was200.0000; the low pass filter was 20000.0000; the number of sample scanswas 128; the collection length was 79.9 seconds; the resolution was4.000; the number of scan points was 8480; the number of FFT points was8192; the laser frequency was 15798.0 cm⁻¹; the interferogram peakposition was 4096; the apodization was triangular; the number ofbackground scans was 128 and the background gain was 1.0.

Differential scanning calorimetric analysis of samples was conducted inthe following manner. A T.A. Instruments Thermal Analyzer 3100 withDifferential Scanning Calorimetry module 2910 was used, along withModulated DSC software version 1.1A. The analysis parameters were:Sample weight: 2.28 mg, placed in a covered, uncrimped aluminum pan;Heating rate: room temperature to 150° C. at 5° C./minute under anitrogen purge.

Example 1 Preparation of Amorphous Ritonavir

Form I crystalline polymorph of ritonavir (100 g) was melted at 125° C.by heating Form I. The melt was maintained at a temperature of 125° C.for 3 hours. The melt was rapidly cooled by placing the containerholding the melt into a Dewar flask containing liquid nitrogen. Theresulting glass was ground with a mortar and pestle to provide amorphousritonavir (100 g). Powder X-ray diffraction analysis confirmed that theproduct was amorphous. Differential scanning calorimetric analysisdetermined that the glass transition point was from about 45° C. toabout 49° C. (Measured onset at 45.4° C. and which ends at 49.08° C.,with a midpoint of 48.99° C.).

Example 2 Preparation of Crystalline Ritonavir (Form II)

Amorphous ritonavir (40.0 g) was dissolved in boiling anhydrous ethanol(100 mL). Upon allowing this solution to cool to room temperature, asaturated solution was obtained. After standing overnight at roomtemperature, the resulting solid was isolated from the mixture byfiltration and was air dried to provide Form II (approximately 24.0 g).

Example 3 Preparation of(2S)-N-((1S)-1-Benzyl-2-((4S,5S)-4-benzyl-2-oxo-1,3-oxazolidin-5-yl)ethyl)-2-((((2-isopropyl-1,3-thiazol-4-yl)methyl)amino)carbonyl)amino)-3-methylbutanamideExample 3a Preparation of(4S,5S)-5-((2S)-2-t-butyloxycarbonylamino-3-phenylpropyl)-4-benzyl-1,3-oxazolidin-2-one

(2S,3S,5S)-2-Amino-3-hydroxy-5-t-butyloxycarbonylamino-1,6-diphenylhexanesuccinate salt (30 g, 63 mmol; U.S. Pat. No. 5,654,466),((5-thiazolyl)methyl)-(4-nitrophenyl)carbonate hydrochloride (22.2 g;U.S. Pat. No. 5,597,926) and sodium bicarbonate (16.2 g) were mixed with300 mL of water and 300 mL of ethyl acetate and the mixture was stirredat room temperature for about 30 minutes. The organic layer was thenseparated and heated at about 60° C. for 12 hours, and then stirred at20-25° C. for 6 hours. 3 mL of ammonium hydroxide (29% ammonia in water)was added and the mixture stirred for 1.5 hours. The resulting mixturewas washed with 4×200 mL of 10% aqueous potassium carbonate and theorganic layer was separated and evaporated under vacuum to provide anoil. The oil was suspended in about 250 mL of heptane. The heptane wasevaporated under vacuum to provide a yellow solid. The yellow solid wasdissolved in 300 mL of THF and 25 mL of 10% aqueous sodium hydroxide wasadded. After stirring for about 3 hours, the mixture was adjusted to pH7 by addition of 4N HCl (about 16 mL). The THF was evaporated undervacuum to leave an aqueous residue, to which was added 300 mL ofdistilled water. After stirring this mixture, a fine suspension ofsolids resulted. The solid was collected by filtration and the filteredsolid was washed with water (1400 mL) in several portions, resulting inthe desired product.

Example 3b Preparation of(4S,5S)-5-((2S)-2-amino-3-phenylpropyl)-4-benzyl-1,3-oxazolidin-2-one

The crude, wet product of Example 3a was slurried in 1N HCl (192 mL) andthe slurry was heated to 70° C. with stirring. After 1 hour, THF (100mL) was added and stirring at 65° C. was continued for 4 hours. Themixture was then allowed to cool to 20-25° C. and was stirred overnightat 20-25° C. The THF was removed by evaporation under vacuum and theresulting aqueous solution was cooled to about 5° C., causing someprecipitation to occur. The aqueous mixture was adjusted to pH 7 byaddition of 50% aqueous sodium hydroxide (about 18.3 g). The resultingmixture was extracted with ethyl acetate (2×100 mL) at about 15° C. Thecombined organic extracts were washed with 100 mL of brine and theorganic layer was separated and stirred with sodium sulfate (5 g) andDarco G-60 (3 g). This mixture was warmed on a hot plate for 1 hour at45° C. The hot mixture was then filtered through a bed of diatomaceousearth and the filter pad was washed with ethyl acetate (100 mL). Thefiltrate was evaporated under vacuum to provide an oil. The oil wasredissolved in methylene chloride (300 mL) and the solvent wasevaporated under vacuum. The resulting oil was dried at room temperatureunder vacuum to provide the desired product (18.4 g) as a glassy syrup.

Example 3c Preparation of(2S)-N-((1S)-1-Benzyl-2-((4S,5S)-4-benzyl-2-oxo-1,3-oxazolidin-5-yl)ethyl)-2-((((2-isopropyl-1,3-thiazol-4-yl)methyl)amino)carbonyl)amino)-3-methylbutanamide

N-((N-Methyl-N((2-isopropyl-4-thiazolyl)methyl)amino)carbonyl)-L-valine(10.6 g, 33.9 mmol; U.S. Pat. No. 5,539,122 and International PatentApplication No. WO98/00410), the product of Example 3b (10.0 g, 32.2mmol) and 1-hydroxybenzotriazole (5.2 g, 34 mmol) were dissolved in THF(200 mL). 1,3-dicylclohexylcarbodiimide (DCC, 7.0 g, 34 mmol) was thenadded to the THF mixture and the mixture was stirred at 22° C. for 4hours. Citric acid (25 mL of 10% aqueous solution) was added andstirring continued for 30 minutes. The THF was then evaporated undervacuum. The residue was dissolved in ethyl acetate (250 mL) and washedwith 10% citric acid solution (175 mL). NaCl (5 g) was added toaccelerate the separation of the layers. The organic layer wassequentially washed with 10% aq. sodium carbonate (2×200 mL) and water(200 mL). The organic layer was then dried over sodium sulfate (20 g),filtered and evaporated under vacuum. The resulting product (20.7 g of afoam) was dissolved in hot ethyl acetate (150 mL) and then heptane (75mL) was added. Upon cooling, another 75 mL of heptane was added and themixture was heated to reflux. Upon cooling to room temperature, noprecipitate formed. The solvents were evaporated under vacuum and theresidue was redissolved in a mixture of 200 mL ethyl acetate/100 mLheptane. The small amount of undissolved solid was removed byfiltration. The filtrate was evaporated under vacuum and the residue wasdissolved in a mixture of 100 mL ethyl acetate/50 mL heptane, giving aclear solution. The solution was cooled to −10° C. and a whiteprecipitate formed. The mixture was allowed to sit at −15° C. for 24hours. The resulting solid was collected by filtration, washed with 1:1ethyl acetate/heptane (2×24 mL) and dried in a vacuum oven at 55° C. toprovide the desired product as a beige solid (16.4 g).

Example 4 Preparation of Crystalline Ritonavir (Form II)

To a solution of 1.595 g of ritonavir Form I in 10 mL of 200 proofethanol was added approximately 50 micrograms of the product of Example3c. This mixture was allowed to stand at about 5° C. for 24 hours. Theresulting crystals were isolated by filtration through 0.45 micron nylonfilter and air dried to provide ritonavir Form II.

Example 5 Alternative Preparation of Crystalline Ritonavir (Form II)

Ethyl acetate (6.0 L/kg of ritonavir) was added to ritonavir (Form I ora mixture of Form I and Form II) in a reaction vessel. The mixture wasstirred and heated to 70° C. until all solids were dissolved. Thesolution was filtered (utilizing a centrifuge pump and 5×20 inchcartridge filters having a porosity of 1.2 microns) and the filtrate wasallowed to cool to 52° C. at a rate of 2-10° C./hour. To this solutionwas added ritonavir Form II seed crystals (about 1.25 g of Form II seedcrystals/kg of ritonavir) and the mixture was stirred at 52° C. for notless than 1 hour at an agitation rate of 15 RPM. The mixture was thenallowed to cool to 40° C. at a rate of 10° C./hour. Heptane (2.8 L/kg ofritonavir) was added at a rate of 7 L/minute with mixing. The mixturewas allowed to cool to 25° C. at a rate of 10° C./hour with mixing. Thenthe mixture was stirred for not less than 12 hours at 25° C. The productwas isolated by filtration using a Heinkel type centrifuge (run timeapproximately 16 hours). The product was dried at 55° C. under vacuum(50 mm Hg) for 16-25 hours to provide ritonavir crystal Form II.

Example 6 Preparation of Amorphous Ritonavir

Ritonavir Form I (40 g) was dissolved in methylene chloride (60 mL).This solution was slowly added over 15 minutes to a round bottom flaskequipped with an overhead stirrer and containing hexanes (3.5 L). Theresulting slurry was allowed to stir for 10 minutes. The precipitate wasfiltered and dried at room temperature in a vacuum oven to provideamorphous ritonavir (40 g).

Example 7 Preparation of Amorphous Ritonavir

Ritonavir Form I (5 g) was dissolved in methanol (8 mL). This solutionwas slowly added to a round bottom flask equipped with an overheadstirrer and containing distilled water (2 L), while maintaining theinternal temperature near 0° C. The resulting solid was filtered to givea sticky solid which was dried in a vacuum oven to give amorphousritonavir (2.5 g).

Example 8 Comparative Solubilities

Solubility experiments were performed for ritonavir Form I and Form IIin various formulation mediums. Data is provided in FIGS. 3-7.

Tables 1 and 2 provided hereinbelow illustrate the pharmaceuticalcomposition without water. Examples 9 and 10 illustrate thepharmaceutical composition with water.

TABLE 1 Composition of Formulations T-1 and T-2. T-1 T-2 Components mg/gmg/cap mg/g mg/cap Ritonavir 200.0 200.0 200.0 200.0 Alcohol,dehydrated, USP 100.0 100.0 100.0 100.0 Oleic acid, NF 650.0 650.0 600.0600.0 Polyoxyl 35 Castor Oil 50.0 50.0 100.0 100.0 (Cremophor EL ®) BHT0.01 0.01 0.01 0.01

TABLE 2 Composition of Formulation T-1B. T-1 B Components mg/g mg/capRitonavir 200.0 200.0 Alcohol, dehydrated, USP 120.0 120.0 Oleic acid,NF 619.5 619.5 Polyoxyl 35 Castor Oil 60.0 60.0 (Cremophor EL ®) BHT 0.50.5

Example 9 Preparation of Norvir® (100 mg) Soft Gelatin Capsules

The following protocol is employed in the preparation of 1000 softgelatin capsules:

Scale Amount (mg/capsule) Name (g) Q.S. Nitrogen, N.F. Q.S. 118.0Ethanol, dehydrated, USP, 200 Proof 118.0 2.0 Ethanol, dehydrated, USP,200 Proof 2.0 0.25 Butylated Hydroxytoluene, NF 0.25 704.75 Oleic Acid,NF 704.75 100.0 Ritonavir 100.0 10.0 Water, purified, USP (distilled)10.0 60.0 Polyoxyl 35 Castor Oil, NF 60.0 5.000 Oleic Acid, NF 5.000

A mixing tank and suitable container are purged with nitrogen. 118.0 gof ethanol is weighed, blanketed with nitrogen, and held for later use.The second aliquot of ethanol (2 g) is then weighed, and mixed with 0.25g of butylated hydroxytoluene until clear. The mixture is blanketed withnitrogen and held. The main mixing tank is heated to 28° C. (not toexceed 30° C.). 704.75 g of oleic acid is then charged into the mixingtank. 100.0 g of ritonavir is then added to the oleic acid with mixing.The ethanol/butylated hydroxytoluene is then added to the mixing tank,followed by the 118.0 g of ethanol measured previously, and mixed for atleast 10 minutes. 10 g of water is then charged into the tank and mixeduntil the solution is clear (for not less than 30 minutes). The sides ofthe vessel are scraped for ritonavir, and mixed for not less than anadditional 30 minutes. 60.0 g of Polyoxyl 35 castor oil is charged intothe tank and mixed until uniform. The solution is stored at 2-8° C.until encapsulation. 1.0 g of the solution is filled into each softgelatin capsule (die: 18 oblong [18BE]; gel: 005L2DDXHB-EP; gel dyes:white 920P). The soft gelatin capsules are then dried, and stored at2-8° C.

Example 10 Preparation of ABT-378/Norvir® (133.3/33.3 mg) Soft GelatinCapsules

The following protocol is employed in the preparation of 1000 softgelatin capsules:

Scale Amount (mg/capsule) Name (g) Q.S. Nitrogen, N.F. Q.S. 578.6 OleicAcid, NF 578.6 33.3 Ritonavir 33.3 64.1 Propylene Glycol, USP 64.1 4.3Water, purified, USP (distilled) 4.3 133.3 ABT-378 133.3 10.0 OleicAcid, NF 10.0 21.4 Polyoxyl 35 Castor Oil, NF 21.4 10.0 Oleic Acid, NF10.0

A mixing tank and suitable container are purged with nitrogen. 578.6 gof oleic acid is then charged into the mixing tank. The mixing tank isheated to 28° C. (not to exceed 31° C.) and mixing is started. 33.3 g ofritonavir is then added to the oleic acid with mixing. The propyleneglycol and water are added to the mixing tank, and mixing is continueduntil the solution is clear. 133.3 g of ABT-378 is then added into themixing tank, and mixing is continued. 10 g of oleic acid is then chargedinto the tank and mixed until the solution is clear. 21.4 g of polyoxy35 Castor Oil, NF is added to the mixing tank, and mixing is continued,followed by the addition of 10 g of Oleic Acid. NF. A sample iscollected, and the solution is stored at 2-8° C. until encapsulation.0.855 (+/1 3%) g of the solution is filled into each soft gelatincapsule (die: 12BF; gel: L1.25DDXHBHM-EP; gel dye: Orange 419T-EP). Thesoft gelatin capsules are then inspected and cleaned, and stored at 2-8°C.

Example 11 Protocol for Oral Bioavailability

Dogs (beagle dogs, mixed sexes, weighing 7-14 kg) were fasted overnightprior to dosing, but were permitted water ad libitum. Each dog receiveda 100 μg/kg subcutaneous dose of histamine approximately 30 minutesprior to dosing. Each dog received a single dosage form corresponding toa 5 mg/kg dose of the drug. The dose was followed by approximately 10milliliters of water. Blood samples were obtained from each animal priorto dosing and 0.25, 0.5, 1.0, 1.5, 2, 3, 4, 6, 8, 10, and 12 hours afterdrug administration. The plasma was separated from the red cells bycentrifugation and frozen (−30° C.) until analysis. Concentrations ofparent drug were determined by reverse phase HPLC with low wavelength UVdetection following liquid-liquid extraction of the plasma samples. Theparent drug area under the curve was calculated by the trapezoidalmethod over the time course of the study. The absolute bioavailabilityof each test composition was calculated by comparing the area under thecurve after oral dosing to that obtained from a single intravenous dose.Each capsule or capsule composition was evaluated in a group containingat least six dogs; the values reported are averages for each group ofdogs.

1. A process of preparing a pharmaceutical composition which includes asolution, said process comprising filling said solution into a capsule,wherein said solution comprises: (a) ritonavir; (b) a pharmaceuticallyacceptable medium and/or long chain fatty acid, or a mixture ofpharmaceutically acceptable medium and/or long chain fatty acids, in anamount of from 30% to 75% by weight of said solution; (c) water in anamount of from 0.4% to 3.5% by weight of said solution; and, optionally,(d) a pharmaceutically acceptable surfactant.
 2. The process accordingto claim 1, wherein said solution further comprise a pharmaceuticallyacceptable alcohol.
 3. The process according to claim 1, wherein saidsolution further comprises ethanol or propylene glycol in an amount offrom 1% to 15% by weight of said solution.
 4. The process according toclaim 3, wherein said solution comprises said pharmaceuticallyacceptable surfactant in an amount of from 2% to 20% by weight of saidsolution.
 5. The process according to claim 3, wherein saidpharmaceutically acceptable medium or long chain fatty acid is amono-unsaturated C₁₆-C₂₀ fatty acid which is liquid at room temperature,and said mixture of pharmaceutically acceptable medium and/or long chainfatty acids is a mixture of mono-unsaturated C₁₆-C₂₀ fatty acids whichare liquids at room temperature.
 6. The process according to claim 3,wherein said pharmaceutically acceptable medium and/or long chain fattyacid, and said mixture of pharmaceutically acceptable medium and/or longchain fatty acids, comprise oleic acid.
 7. The process according toclaim 1, wherein said solution comprises: (a) ritonavir in an amount offrom 1% to 30% by weight of said solution; (b) oleic acid in an amountof from 30% to 75% by weight of said solution; and (c) polyoxyl 35castor oil in an amount of from 0% to 20% by weight of said solution. 8.The process according to claim 7, wherein said solution furthercomprises ethanol in an amount of from 1% to 15% by weight of saidsolution.
 9. The process of according to claim 8, wherein said solutioncomprises polyoxyl 35 castor oil in an amount of from 2.5% to 10% byweight of said solution.
 10. The process according to claim 1, whereinsaid solution comprises: (a) ritonavir and(2S,3S,5S)-2-(2,6-dimethylphenoxyacetyl)-amino-3-hydroxy-5-(2S-(1-tetrahydropyrimid-2-onyl)-3-methyl-butanoyl)amino-1,6-diphenylhexanein an amount from 1% to 45% by weight of said solution; (b) oleic acidin an amount of from 30% to 75% by weight of said solution; and (c)polyoxyl 35 castor oil in an amount of from 0% to 20% by weight of saidsolution.
 11. The process according to claim 10, wherein said solutioncomprises propylene glycol in an amount of from 1% to 15% by weight ofsaid solution.
 12. The process according to claim 11, wherein saidsolution comprises polyoxyl 35 castor oil in an amount of from 2.5% to10% by weight of said solution.
 13. The process according to claim 1,wherein said capsule is a hard gelatin capsule or a soft gelatincapsule.
 14. A method for improving the pharmacokinetics of a drug whichis metabolized by cytochrome P450 monooxygenease, said method comprisingadministering to a human in need thereof a combination of said drug anda pharmaceutical composition prepared according to the process ofclaim
 1. 15. A process of preparing a pharmaceutical composition whichincludes a solution, said process comprising filling said solution intoa capsule, wherein said solution comprises: (a) ritonavir in an amountof 10% by weight of said solution; (b) oleic acid in an amount of from70% to 75% by weight of said solution; (c) ethanol in an amount of from3% to 12% by weight of said solution; (d) water in an amount of from0.4% to 1.5% by weight of said solution; and (e) polyoxyl 35 castor oilin an amount of 6% by weight of said solution.
 16. A method forimproving the pharmacokinetics of a drug which is metabolized bycytochrome P450 monooxygenease, said method comprising administering toa human in need thereof a combination of said drug and a pharmaceuticalcomposition prepared according to the process of claim
 15. 17. Themethod of claim 16, wherein said drug is an immunosuppressant, achemotherapeutic agent, an antibiotic, or an HIV protease inhibitor. 18.An unencapsulated solution comprising: (a) ritonavir; (b) apharmaceutically acceptable medium and/or long chain fatty acid, or amixture of pharmaceutically acceptable medium and/or long chain fattyacids, in an amount of from 30% to 75% by weight of said solution; (c)water in an amount of from 0.4% to 3.5% by weight of said solution; and,optionally, (d) a pharmaceutically acceptable surfactant.
 19. Theunencapsulated solution of claim 18, further comprising apharmaceutically acceptable alcohol.
 20. The unencapsulated solution ofclaim 18, further comprising ethanol or propylene glycol in an amount offrom 1% to 15% by weight of said solution.